This study confirmed that the recombinant humanized collagen type III (rhCol III) produced by Shanxi Jinbo Bio-pharmaceutical Co., Ltd. conforms to the theoretical amino acid sequence (excluding the non-human collagen amino acid sequence) and has a triple helix structure. It can obviously promote the adhesion, proliferation and migration of human skin fibroblasts. This study also found that rhCol III could express the components of extracellular matrix (ECM) and it will help to further study the mechanism of the interaction between rhCol and cells and further verify its effectiveness in tissue regeneration.